The components of the corneal cell surface and its extracellular matrix play a very important role in corneal metabolism throughout the normal process of growth and aging. Many abnormal changes of the cornea can probably be directly traced to the metabolism of the cell surface and the extracellular matrix. The overall objectives of this research are to obtain specific monoclonal immunological reagents against certain antigens in human and mouse corneal tissues and cells in culture, and to use these reagents to define similarities and differences between the cornea and other tissues. Monoclonal antibody and specific recombinant DNA probes will be used to determine whether certain antigens are being expressed in various human-mouse somatic cell hybrids or mouse-chinese hamster hybrids. By knowing the chromosome constitution of the various hybrids, the presence or absence of the antigen or recombinant probe will determine whether the specific chromosome is present in the hybrid cells, thus localizing the gene to its chromosome. The methodologies to be used in the majority of the proposed studies involve hybridoma and recombinant DNA technology. Mice are injected with protein or live cells to generate an antibody response. After a certain time, or after several injections, the spleen is removed from the animal and the lymphocytes are hybridized with mutant myeloma cells to produce a hybridoma cell. The cells are cloned and grown to large quantities and monoclonal antibody is purified from the culture medium. Characterization of the antibodies and determination of the specific antigen to which the antibody reacts involves immunoprecipitation, immunoelectrophoresis, PAGE, isoelectric focusing, column chromatography, immunostaining of cells and tissues, and flow cytometry. Monoclonal antibody will be used as a probe to isolate the gene coding for each of the corneal antigens. Other methodologies to be used involve somatic cell hybridization, and chromosome identification. Our studies may lead to insights into the dynamic changes occurring in the cornea throughout the process of normal growth and those changes occuring in disease states and following trauma and may provide answers to many unknown questions involving pathogenesis of the cornea and in the senscent state.